Journal: Bioactive Materials

Article Title: “Slow walk” mimetic tensile loading maintains human meniscus tissue resident progenitor cells homeostasis in photocrosslinked gelatin hydrogel

doi: 10.1016/j.bioactmat.2023.01.025

Figure Lengend Snippet: Design and application of GelMA-based mechanical actuator to provide biomimetic cyclic tensile loading to hydrogel-encapsulated hMeSPCs. (a) Diagrammatic representation of tensile loading in the native meniscal resident cells in the load-bearing joint, specifically showing joint load-bearing of meniscus tissue at a "slow walk" speed. (b) Elastic moduli of the ECM and PCM of various regions of the native porcine meniscus at the microscopic level, evaluated by AFM; data adopted from Ref. . (c) Young's modulus values of 10% and 15% w/v GelMA with 30%, 60%, and 90% DS. *, p < 0.05; **, p < 0.01; ***, p < 0.001. Data are mean ± SD, n ≥ 6. (d) Operating principle of the actuator. Anchoring point: big block of GelMA with anchor to fix the whole hydrogel construct in the cell culture plate in a certain position; Loading arms: GelMA hydrogel with/without cells can be loaded in the loading arms, and the magnetic beads are loaded in the end of the loading arms; S0, loading arms fixation point; S1, the end position of loading arms without the influence of magnetic field; S2, hydrogel elongation position when the magnetic field is applied; a photo of the actuator can be found in . (e) Picture of the GelMA constructs: GelMA hydrogel elongation in response to external magnetic stimuli to provide cyclic tensile loading on the loading arms. (f) Swelling ratio of the GelMA hydrogel (10% w/v, 60% DS) under static and loading conditions at days 0, 5, 10, and 15. (g) Harvesting strategy for hMeSPCs.

Article Snippet: With the 3D GelMA hydrogel cyclic loading cell culture system, we found that a “slow walk” biomimetic cyclic loading regimen (10% tensile strain, 0.5 Hz, 1 h/day, up to 15 days) significantly increased (i) hMeSPC differentiation, ( ii ) fibrocartilage-like ECM deposition, and ( iii ) the mechanobiological response of 3D encapsulated hMeSPCs and their interactions with GelMA hydrogel.

Techniques: Blocking Assay, Construct, Cell Culture, Magnetic Beads

Journal: Bioactive Materials

Article Title: “Slow walk” mimetic tensile loading maintains human meniscus tissue resident progenitor cells homeostasis in photocrosslinked gelatin hydrogel

doi: 10.1016/j.bioactmat.2023.01.025

Figure Lengend Snippet: Cell viability, cell morphology, and mechanical property of hMeSPC-GelMA constructs after mechanical loading. (a) Mechanical loading regimen for GelMA hydrogel, consisting of 10% elongation under cyclic loading. S0, fixation point; S1, starting point; S2, 10% hydrogel elongation, at which point the tensile load drops off and the hydrogel starts to restore to the relaxation point, S3; S4, maximum elongation point, where the loading arms are maximally elongated in response to magnetic force at this point. (b) LIVE/DEAD staining of hMeSPCs encapsulated in GelMA hydrogel (10% w/v, 60% DS) after static culture and tensile loading at days 0, 5, 10, 15. Left panel: green, live cells; right panel: red, dead cells. Scale bar = 100 μm. (c) Flow cytometric analysis of PI stained hMeSPCs released after 2D culture ( Ctrl ), and 3D cultures maintained under static ( Static ), and tensile loading ( Loaded , 10% elongation, 0.5 Hz, 1 h/day) conditions at day 15. The % of dead cells are estimated based on the relative number of PI positive cells in the total population. 3 batches (n = 9 biological donors) were examined with 3 technical repeats. Representative data from 1 batch is shown. (d) Representative images of cytoskeletal morphology (F-actin, phalloidin-iFluor 555 staining, red; nuclei, DAPI staining, blue). (e) Cross-sectional cell shape aspect ratio of the encapsulated hMeSPCs under static and loaded conditions at days 0, 5, 10, 15. Data were analyzed by ANOVA; ***, p < 0.001 between static and loaded groups at days 10 and 15. Data are mean ± SD; n = 8 random views from high magnification field (HMF) per group. (f) Quantification of protrusions in hMeSPCs under static and loaded conditions at days 0, 5, 10, and 15. N = the number of counted cells in hydrogel. Data are mean ± SD, analyzed by ANOVA; ** , p < 0.01 and **** , p < 0.0001 between static and loaded groups at days 10 and 15, respectively. (g) Representative pictures of stress-strain curves of GelMA constructs with or without encapsulated hMeSPCs, maintained under static and loaded conditions at days 0, 5, 10, 15. Data were collected from n = 3 batches with total of 9 biological donors; each experiment was performed with triplicates as technical repeats. The X axis is strain, and the number represent length change (mm)/original length (mm), and the Y axis is stress (MPa). (h) Young's modulus values of GelMA constructs with or without encapsulated hMeSPCs, maintained under static and loaded conditions at days 0, 5, 10, 15. Data were collected from n = 3 batches with total of 9 biological donors; each experiment was performed with triplicates as technical repeats. Two-way ANOVA with post-hoc was used to study the difference between groups; Data are mean ± SD. GelMA only-Loaded groups vs GelMA only-Static groups: *** , p < 0.001 and **** , p < 0.0001; GelMA + cell-Loaded groups vs GelMA + cell-Static groups: ^ , p < 0.05; GelMA + cell-Static groups vs GelMA only-Static groups: #### , p < 0.0001; GelMA + cell-Loaded groups vs GelMA only-Loaded groups: & , p < 0.05 and && , p < 0.01. (i) Maximum elongation (S3–S4) of GelMA hydrogel cultures at days 5, 10, and 15 after loading, normalized to initial length of GelMA-hydrogel arms (S0–S3). Two-way ANOVA: *, p < 0.05 between day 15 and day 0 in GelMA only group; and # , p < 0.05 between day 15 and day 5 in GelMA only group. Data were collected from n = 3 batches with a total of 9 biological donors; each experiment was performed with triplicates as technical repeats.

Article Snippet: With the 3D GelMA hydrogel cyclic loading cell culture system, we found that a “slow walk” biomimetic cyclic loading regimen (10% tensile strain, 0.5 Hz, 1 h/day, up to 15 days) significantly increased (i) hMeSPC differentiation, ( ii ) fibrocartilage-like ECM deposition, and ( iii ) the mechanobiological response of 3D encapsulated hMeSPCs and their interactions with GelMA hydrogel.

Techniques: Construct, Staining

Journal: Bioactive Materials

Article Title: “Slow walk” mimetic tensile loading maintains human meniscus tissue resident progenitor cells homeostasis in photocrosslinked gelatin hydrogel

doi: 10.1016/j.bioactmat.2023.01.025

Figure Lengend Snippet: Biomimetic tensile loading of hMeSPCs enhanced meniscus-like ECM generation and deposition. (a & b) Safranin O staining of the histologic sections of hMeSPCs-GelMA constructs cultured with (loaded) or without (static) biomimetic loading at 0, 5, 10, and 15 days ( a, blue arrowheads indicating positively stained areas), and ( b ) quantification of Safranin O-stained areas. (c & d) Immunostaining for Col I deposition ( c , red arrowheads) in the hMeSPCs-GelMA constructs cultured with or without biomimetic loading at days 0, 5, 10, 15, and ( d ) quantification of Col I positive staining area. (e) Immunostaining for Col II deposition ( e , red arrowheads) by hMeSPCs with or without biomimetic loading at days 0, 5, 10, 15, and (f) quantification of Col II positive staining area. Histological analysis of each hydrogel construct shown above was performed at three different regions of interests (ROIs) of each culture under 10× magnification of light microscopy. The positive staining area within GelMA hydrogel structure was quantified using Image J (NIH, US). The experiment was repeated three times with cells from B1, B2 and B3, separately, with 3 technical repeats in each group. Two-way ANOVA with post-hoc between groups; *, p < 0.05; **, p < 0.01; ***, p < 0.001. Data are mean ± SD, scale bar = 100 μm. (g) Ratio of Col II to Col I immunostained areas in hMeSPCs-GelMA constructs cultured with (loaded) or without (static) biomimetic loading. Two-way ANOVA with post-hoc between groups; *, p < 0.05. Data are mean ± SD, n = 3. (h) Diagrammatic representation of the ECM content in the inner and outer meniscus tissue. Static , Loaded: as described in .

Article Snippet: With the 3D GelMA hydrogel cyclic loading cell culture system, we found that a “slow walk” biomimetic cyclic loading regimen (10% tensile strain, 0.5 Hz, 1 h/day, up to 15 days) significantly increased (i) hMeSPC differentiation, ( ii ) fibrocartilage-like ECM deposition, and ( iii ) the mechanobiological response of 3D encapsulated hMeSPCs and their interactions with GelMA hydrogel.

Techniques: Staining, Construct, Cell Culture, Immunostaining, Light Microscopy

Journal: Bioactive Materials

Article Title: “Slow walk” mimetic tensile loading maintains human meniscus tissue resident progenitor cells homeostasis in photocrosslinked gelatin hydrogel

doi: 10.1016/j.bioactmat.2023.01.025

Figure Lengend Snippet: Degradation of GFT-GelMA hydrogel constructs with or without encapsulated hMeSPCs under intermittent tensile loading. (a) Picrosirius Red staining (collagen stain: pink for thin fibers and red for thick fibers) of histologic sections of the hMeSPCs-GelMA constructs under static and loaded conditions at days 0, 5, 10, and 15. Representative micrographs from 3 batches of cultures. Scale bar = 100 μm. (b) Histomorphometric analysis of porosity as a function of culture time, in terms of average pore area (diameter 2 , d 2 ) and number of pores in the indicated ranges of pore area. (c) Porosity of GelMA constructs with and without encapsulated hMeSPCs cultured under static and loaded conditions at days 0, 5, 10, and 15. Porosity is calculated as total area of pores (number × π.d 2 /4) expressed as a percentage of total area of the microscopic field. In ( b ) and ( c ), data were collected from 3 batches of cultures consisting of cells derived from 9 biological donors, with triplicates as technical repeats. In ( c ), Two-way ANOVA with post-hoc was used to compare between groups. Data are mean ± SD. GelMA only-Loaded groups vs GelMA only-Static groups: ** , p < 0.01; GelMA + cell-Static groups vs GelMA only-Static groups: ## , p < 0.01, ### , p < 0.001; GelMA + cell-Loaded groups vs GelMA only-Loaded groups: & , p < 0.05. (d) Representative GFT fluorescence (left panels) and corresponding heatmap intensity of GFT loss (FIH, right panels) images of GFT-GelMA hydrogel constructs with or without encapsulated hMeSPCs cultured under static and loaded conditions at days 0, 5, 10, and 15. Scale bar = 100 μm. Representative micrographs from 3 batches of cultures consisting of cells derived from 9 biological donors. (e) Daily release of fluorescent soluble products derived from degraded GFT-GelMA constructs with or without encapsulated hMeSPCs cultured under static and loaded conditions. GFT-GelMA (60% DS) was used at 10% w/v and hMeSPCs were seeded at 2.5 × 10 5 cells per 50 μl GelMA. Data (arbitrary units, mean ± SD) were collected from 3 batches of cultures consisting of cells derived from 9 biological donors, with triplicates as technical repeats. Two-way ANOVA with post-hoc between groups. GelMA + cell-Static groups vs GelMA only-Static groups: * , p < 0.05, ** , p < 0.01, *** , p < 0.001; GelMA + cell-Loaded groups vs GelMA only-Loaded groups: # , p < 0.05, ## , p < 0.01, ### , p < 0.001. (f) Expression of ECM dynamic-related genes regulated by intermittent tensile stimulation (loaded vs static; red, up-regulated; blue, down-regulated), and classified by biological function, based on transcriptome analysis described in . (g) Protein-protein interaction network of the corresponding regulated genes. Data were included and analyzed from RNA-seq data intersection of B1 and B2. Static , Loaded: as described in .

Article Snippet: With the 3D GelMA hydrogel cyclic loading cell culture system, we found that a “slow walk” biomimetic cyclic loading regimen (10% tensile strain, 0.5 Hz, 1 h/day, up to 15 days) significantly increased (i) hMeSPC differentiation, ( ii ) fibrocartilage-like ECM deposition, and ( iii ) the mechanobiological response of 3D encapsulated hMeSPCs and their interactions with GelMA hydrogel.

Techniques: Construct, Staining, Cell Culture, Derivative Assay, Fluorescence, Expressing, RNA Sequencing

Journal: Bioactive Materials

Article Title: “Slow walk” mimetic tensile loading maintains human meniscus tissue resident progenitor cells homeostasis in photocrosslinked gelatin hydrogel

doi: 10.1016/j.bioactmat.2023.01.025

Figure Lengend Snippet: Diagrammatic representation of ECM dynamics in hMeSPC-GelMA hydrogel constructs cultured under biomimetic tensile loading conditions. Red, hydrogel construct retention; Pink, GelMA degradation; and green, ECM deposition.

Article Snippet: With the 3D GelMA hydrogel cyclic loading cell culture system, we found that a “slow walk” biomimetic cyclic loading regimen (10% tensile strain, 0.5 Hz, 1 h/day, up to 15 days) significantly increased (i) hMeSPC differentiation, ( ii ) fibrocartilage-like ECM deposition, and ( iii ) the mechanobiological response of 3D encapsulated hMeSPCs and their interactions with GelMA hydrogel.

Techniques: Construct, Cell Culture

Journal: Bioengineering

Article Title: Bioengineered Approaches for Esophageal Regeneration: Advancing Esophageal Cancer Therapy

doi: 10.3390/bioengineering12050479

Figure Lengend Snippet: Trends in publication counts from 1990 to 2023 on esophageal regenerative and reconstructive approaches, including esophageal reconstruction surgery, esophageal transplantation surgery, esophageal tissue engineering, and bioprinting. Data were compiled and analyzed by the author using Google Scholar search results.

Article Snippet: Square , ~3 × 5 mm 2 , Patch , 3D bioprinting (GelMA, SFMA, Fe 3 O 4 , BMSC) , 9 days , Hydrogel scaffold supports cell growth and differentiation, aligning BMSCs into SMCs to create a transplantable biomimetic muscle construct It effectively restores smooth muscle structure by enhancing SMC alignment and ECM remodeling , [ ] .

Techniques: Transplantation Assay

Journal: Biomaterials

Article Title: Dental Cell Sheet Biomimetic Tooth Bud Model

doi: 10.1016/j.biomaterials.2016.08.024

Figure Lengend Snippet: A. DE and DM cells were seeded on thermo-responsive plates and cultured in normal DE and DM media, respectively, for 14 days. DE and DM CSs were detached by temperature reduction (20ºC) and layered over GelMA constructs to create experimental 3D tooth bud constructs (CSG = DE and DM CSs layered over dental cells encapsulated in GelMA; G = GelMA alone). For in vivo analyses, replicate constructs were cultured in osteogenic media for 4 days and implanted subcutaneously onto the backs of the rats. B. Bioengineered 3D CS - GelMA tooth bud model. The bottom layer mimics the pulp organ (5% GelMA encapsulating DM cells) and the top layer mimics the enamel organ (3% GelMA encapsulating DE cells). The DE and DM CS layers mimic polarized DE-DM cell layers normally observed in developing teeth. C. Steps used to prepare the constructs. DM cells (3×107 cells/ml) were re-suspended in 100 μL of 5% GelMA and photo-crosslinked. DM and DE cell sheets were layered over the polymerized DM 5% GelMA. DE cells (3×107 cells/ml) re-suspended in 100 μL 3% GelMA and 100 μL, layered over construct and photo-crosslinked.

Article Snippet: Moreover, to create a biomimetic tooth bud, and to increase the DM-DM, DE-DE, and DE-DM cell interactions observed in natural tooth bud development, DE-DM dental cell sheets were sandwiched between biomimetic 3D GelMA enamel and pulp organs.

Techniques: Cell Culture, Construct, In Vivo

Journal: Biomaterials

Article Title: Dental Cell Sheet Biomimetic Tooth Bud Model

doi: 10.1016/j.biomaterials.2016.08.024

Figure Lengend Snippet: H&E images (A, B, C, D) revealed extracellular matrix formation and the morphology of the DE and DM cell sheets within the bilayer GelMA constructs. The arrows indicate the DE and the DM CSs. Pol images (E, F, G, H) show the organized collagen in the extracellular matrix. IF imaging (I, J, K, L) showed the expression of VM (green) by DM cells and CK18 (red) by the DE cells.

Article Snippet: Moreover, to create a biomimetic tooth bud, and to increase the DM-DM, DE-DE, and DE-DM cell interactions observed in natural tooth bud development, DE-DM dental cell sheets were sandwiched between biomimetic 3D GelMA enamel and pulp organs.

Techniques: Construct, Imaging, Expressing

Journal: Biomaterials

Article Title: Dental Cell Sheet Biomimetic Tooth Bud Model

doi: 10.1016/j.biomaterials.2016.08.024

Figure Lengend Snippet: High magnification H&E images and IHC analyses of multilayered DE DM CSs GelMA constructs cultured in osteogenic media for 24 h and 4 days, stained with FAK, TEN and SYN4. Arrows indicate expression. Cell sheets are identified as epithelial (DE) and mesenchymal (DM). A. H&E stained DE DM CSs GelMA constructs cultured in osteogenic media for 24 h. FAK, TEN and SYN4 staining (B, C and D) were detected in the DM CSs cultured in osteogenic media for 24 h. E. H&E image of DE DM CSs GelMA constructs cultured in osteogenic media for 4 days. F. FAK staining was detected in DE and DM CSs cultured in osteogenic media for 4 days. G. TEN was detected in the DM CSs cultured in osteogenic media for 4 days. H. Faint SYN4 staining was detected in DM CSs cultured in osteogenic media for 4 days. I. No staining was detected in the negative controls. Specific staining was detected on the natural tooth bud (J. FAK, K. TEN and L. SYN4).

Article Snippet: Moreover, to create a biomimetic tooth bud, and to increase the DM-DM, DE-DE, and DE-DM cell interactions observed in natural tooth bud development, DE-DM dental cell sheets were sandwiched between biomimetic 3D GelMA enamel and pulp organs.

Techniques: Construct, Cell Culture, Staining, Expressing

Journal: Biomaterials

Article Title: Dental Cell Sheet Biomimetic Tooth Bud Model

doi: 10.1016/j.biomaterials.2016.08.024

Figure Lengend Snippet: H&E images and IF analyses of multilayered DE DM CSs GelMA constructs cultured in osteogenic media for 24 h and 4 days, stained with SHH, RUNX2 and BMP2 in green, and VM positive DM cells in red. The red staining identifies the DM CSs, while, the absence of red staining identifies the DE cells. Arrows indicate expression. H&E stained DE DM CSs GelMA constructs cultured in osteogenic media for 24 h (A) and 4 days (E). SHH staining was detected in DE and DM CSs after 24 h (B) and 4 days (F). RUNX2 staining was faintly detected at the interface of DE and DM CSs (C), but strongly detected at the second layer of DE CSs after 24 h (inset in the image C), and detected in DE and DM CSs after 4 days (G). BMP2 staining was detected in DE and DM CSs after 24 h (D) and 4 days (H). No staining was detected in the negative controls (I, J and K).

Article Snippet: Moreover, to create a biomimetic tooth bud, and to increase the DM-DM, DE-DE, and DE-DM cell interactions observed in natural tooth bud development, DE-DM dental cell sheets were sandwiched between biomimetic 3D GelMA enamel and pulp organs.

Techniques: Construct, Cell Culture, Staining, Expressing

Journal: Biomaterials

Article Title: Dental Cell Sheet Biomimetic Tooth Bud Model

doi: 10.1016/j.biomaterials.2016.08.024

Figure Lengend Snippet: A. In vivo implanted 3 week constructs at harvest (G is acellular GelMA, CSG is biomimetic 3D CSs GelMA construct). B. Bright field images of an in vivo CSG construct. C. Bright field image of an in vivo acellular GelMA constructs.

Article Snippet: Moreover, to create a biomimetic tooth bud, and to increase the DM-DM, DE-DE, and DE-DM cell interactions observed in natural tooth bud development, DE-DM dental cell sheets were sandwiched between biomimetic 3D GelMA enamel and pulp organs.

Techniques: In Vivo, Construct

Journal: Biomaterials

Article Title: Dental Cell Sheet Biomimetic Tooth Bud Model

doi: 10.1016/j.biomaterials.2016.08.024

Figure Lengend Snippet: A. No mineralized tissue formation was observed in the acellular GelMA constructs (G). B. Mineralized tissue formation was observed in the CSG constructs. C. 3D model of the mineralized tissue. D. Quantification of mineral density (g/cm3) of the CSG constructs. E. Comparison of mineral densities from engineered and natural mineralized tissues (pig spine, trabecular bone, cortical bone and human enamel) [1, 2]. F. Percent volume of mineralized tissue within ranges of mineral density (ROI – region of interest corresponds to the whole mineralized tissue). G. Representation of areas of mineralized tissue within the ranges of mineral densities (white color represents areas within the range). Abbreviations: MD, mineral density.

Article Snippet: Moreover, to create a biomimetic tooth bud, and to increase the DM-DM, DE-DE, and DE-DM cell interactions observed in natural tooth bud development, DE-DM dental cell sheets were sandwiched between biomimetic 3D GelMA enamel and pulp organs.

Techniques: Construct, Comparison

Journal: Biomaterials

Article Title: Dental Cell Sheet Biomimetic Tooth Bud Model

doi: 10.1016/j.biomaterials.2016.08.024

Figure Lengend Snippet: No tissue formation was observed in the acellular GelMA constructs, H&E (A) and Pol (B) images. H&E stained embedded paraffin and sectioned constructs exhibited high cellularity (C, D), extensive extracellular matrix and dentin/bone-like tissue formation at the DM GelMA layer. The dashed line separates the biomimetic pulp organ (DM in the bottom layer) from the biomimetic enamel organ (DE in the top layer). Pol images (E, F) revealed organized collagen formation within the CSG constructs. IF images (G, H) show the expression of VM (green) by DM cells in the biomimetic pulp organ layer, and ECAD (red) by the DE cells in the biomimetic enamel organ of the CSG constructs.

Article Snippet: Moreover, to create a biomimetic tooth bud, and to increase the DM-DM, DE-DE, and DE-DM cell interactions observed in natural tooth bud development, DE-DM dental cell sheets were sandwiched between biomimetic 3D GelMA enamel and pulp organs.

Techniques: Construct, Staining, Expressing

Journal: Journal of tissue engineering and regenerative medicine

Article Title: Developing a biomimetic tooth bud model

doi: 10.1002/term.2246

Figure Lengend Snippet: Comparative elastic moduli of gelatin methacrylate (GelMA) constructs and natural porcine dental tissues. (a) GelMA Gel formulae with corresponding GelMA and photoinitiator concentrations (% w/v). Elastic moduli of (b) unseeded GelMA constructs, (c) porcine dental epithelial (pDE)–porcine dental mesenchymal (pDM) cell-encapsulated GelMA constructs, and (d) natural porcine dental tissues. Dental cell-seeded Gel 3 had similar elastic modulus to that of pDM tissue. Bar graphs represent average ± SD (n = 3). ND, not determined (elastic modulus below detection level). ***p ≤ 0.001; ANOVA followed by Sidak s comparison

Article Snippet: Biomimetic 3D GelMA tooth bud constructs provide a promising model for the eventual development of functional, bioengineered replacement teeth of specified size and shape.

Techniques: Construct, Comparison

Journal: Journal of tissue engineering and regenerative medicine

Article Title: Developing a biomimetic tooth bud model

doi: 10.1002/term.2246

Figure Lengend Snippet: Capillary-like network formation within in vitro-cultured porcine dental mesenchymal (pDM)–human umbilical vein endothelial cells (HUVECs) gelatin methacrylate (GelMA) constructs. (a,b) pDM–HUVEC Gel 3 construct and (c) porcine dental epithelial (pDE)-HUVEC Gel 3 construct. Vascular network formation was observed in pDM–HUVEC GelMA Gel 3 constructs after 4 weeks of in vitro culture (a, arrows). Confocal analyses revealed organized pDM–HUVEC structures (b). No capillary-like formation was observed in pDE–HUVEC constructs (c). Bar: (a,c) 50μm (b) 10 μm.

Article Snippet: Biomimetic 3D GelMA tooth bud constructs provide a promising model for the eventual development of functional, bioengineered replacement teeth of specified size and shape.

Techniques: In Vitro, Cell Culture, Construct

Journal: Journal of tissue engineering and regenerative medicine

Article Title: Developing a biomimetic tooth bud model

doi: 10.1002/term.2246

Figure Lengend Snippet: Parallel in vitro and in vivo bioengineered three-dimensional gelatin methacrylate (GelMA) tooth bud constructs. (a) Schematic of construct fabrication. (b) Experimental timeline. (c–j) Harvested in vivo implanted GelMA tooth bud constructs. Representative bright field images of replicate in vivo GelMA constructs harvested after 3 weeks (c–f) or 6 weeks (g–j) implantation. (c’–j’) Radiographic images of corresponding bright field images indicate mineralized tissue formation (arrows) in 3-week and 6-week constructs. Bar: 2 mm. DE, dental epithelial cell; DM, dental mesenchymal cell; HUVEC, human umbilical vein endothelial cell.

Article Snippet: Biomimetic 3D GelMA tooth bud constructs provide a promising model for the eventual development of functional, bioengineered replacement teeth of specified size and shape.

Techniques: In Vitro, In Vivo, Construct

Journal: Journal of tissue engineering and regenerative medicine

Article Title: Developing a biomimetic tooth bud model

doi: 10.1002/term.2246

Figure Lengend Snippet: Dental cell and human umbilical vein endothelial cell (HUVEC) distribution within in vivo gelatin methacrylate (GelMA) tooth bud constructs. (a–c) Hematoxylin and eosin (H&E) staining revealed high cellularity and the development of bone-like tissue over time. E-cadherin (Ecad)-expressing porcine dental epithelial (pDE) cells (d–f, d’–f’ arrows) and vimentin (VM)-expressing porcine dental mesenchymal (pDM) cells (g–i, g’–i’ arrows) were detected throughout the constructs. CD31-expressing HUVECs were also detected throughoutthe constructs (j–l, j’–l’) and contributed to vascular networks in 3-weekand 6-week in vitro-cultured constructs (k’,l’ arrows ). (d’–l’) Higher magnifications of boxed regions in d–l. Bar: (a–l) 200 μm, (d’–l’) 50 μm.

Article Snippet: Biomimetic 3D GelMA tooth bud constructs provide a promising model for the eventual development of functional, bioengineered replacement teeth of specified size and shape.

Techniques: In Vivo, Construct, Staining, Expressing, In Vitro, Cell Culture

Journal: Journal of tissue engineering and regenerative medicine

Article Title: Developing a biomimetic tooth bud model

doi: 10.1002/term.2246

Figure Lengend Snippet: Dental cell differentiation within in vivo gelatin methacrylate (GelMA) tooth bud constructs. A–i Immunohistochemical analyses of tooth and bone specific markers in 1-, 3- and 6-week in vivo constructs. The odontoblast differentiation marker dentin sialophosphoprotein (DSPP) was detected throughout the constructs at each time-point (a–c, a’–c’). Odontoblast/osteoblast differentiationmarker osteocalcin (OC) expression increased overtime invivo (d–f, d’–f’).Ameloblast differentiationmarker amelogenin (AM) was detected throughout the constructs at all times (g–i, g’–i’). (a’–i’) Higher magnification images of boxed regions in a–i. Bar: (a–i) 200 μm, (a’–i’) 50 μm.

Article Snippet: Biomimetic 3D GelMA tooth bud constructs provide a promising model for the eventual development of functional, bioengineered replacement teeth of specified size and shape.

Techniques: Cell Differentiation, In Vivo, Construct, Immunohistochemical staining, Marker, Expressing

Journal: Journal of tissue engineering and regenerative medicine

Article Title: Developing a biomimetic tooth bud model

doi: 10.1002/term.2246

Figure Lengend Snippet: Schematic of bioengineered neovascular formation in gelatin methacrylate (GelMA) tooth bud constructs. (a) Cross-sectional and (b) longitudinal schematic along with a (c) color-coded key depicting the organization of normal blood vessel, in vitro-cultured GelMA construct capillary network formation, and neovascularization and mineralization of in vivo implanted GelMA constructs. AM, amelogenin; DSPP, dentin sialophosphoprotein; HUVEC, human umbilical vein endothelial cell; OC, osteocalcin; pDE, porcine dental epithelial cell; pDM, porcine dental mesenchymal cell.

Article Snippet: Biomimetic 3D GelMA tooth bud constructs provide a promising model for the eventual development of functional, bioengineered replacement teeth of specified size and shape.

Techniques: Construct, In Vitro, Cell Culture, In Vivo